UbG76V-eGFP-V5His_20aa
(Plasmid #23970)

• Depositing Lab: Nico Dantuma
• Publication: Beskow et al J Mol Biol. 2009 Dec 11. 394(4):732-46.

Backbone

• Vector backbone: EGFP-N1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 3970
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Ub-G76V-eGFP-V5HIS_20aa
• Alt name: Ubiquitin
• Alt name: Ub
• Alt name: Ub-g76v-eGFP
• Insert Size (bp): 990
• Mutation: G76V mutation in Ubiquitin. Ub fused to N-terminus of EGFP. Extended C-terminal peptide sequence after EGFP containing first 20aa of a modified V5HIS tag
• GenBank ID: NM_018955
• Tag / Fusion Protein:
  • eGFP (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Sac1 (not destroyed)
• 3′ cloning site: NotI (not destroyed)
• 5′ sequencing primer: CMV-F

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Please note that this plasmid does not express a functional V5-His tag. This deletion construct was used to determine the minimum peptide length of a C-terminal tag to allow degradation of UbG76V in cells depleted of p97. The translated fusion protein ends with the following sequence: SCRPL ESRGP FEGKP IPNPL*

For more information, see the associated publication.